Gene transfer into CD4+ T lymphocytes:: Green fluorescent protein-engineered, encephalitogenic T cells illuminate brain autoimmune responses
Gene transfer into CD4+ T lymphocytes:: Green fluorescent protein-engineered, encephalitogenic T cells illuminate brain autoimmune responses
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DOI：
10.1038/10567
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发表时间：
1999-07-01
期刊：
影响因子：
82.9
通讯作者：
Wekerle, H
中科院分区：
文献类型：
--
作者：
Flügel, A;Willem, M;Wekerle, H
图1逆转录病毒基因载体及GFP在CD4+ T淋巴细胞中的表达。a，逆转录病毒载体构建pLGFPSN用于转导CD4+ T淋巴细胞。5 ' LTR，长末端重复序列（5 '）驱动转基因的表达；SD和SA，剪接供体和剪接受体序列；猿猴病毒40启动子驱动新霉素耐药基因NEO；3 ' LTR，含有聚腺苷酸化序列的长末端重复序列（3 '）。b， GFP转导后mbp特异性t淋巴细胞的相衬（左）和荧光显微镜（绿色，中间面板）。将细胞铺在载玻片上，用小鼠抗大鼠cd4单克隆抗体染色（右图红色）。用pe标记的山羊抗小鼠血清作为二抗。c，激活依赖性GFP表达CD4+ T淋巴细胞。激活（上）和静止（下）转导和非转导T淋巴细胞的直方图和点阵图。点图显示gfp转导的T细胞CD4染色。图中显示了15个以上的代表性细胞系。
Fig. 1 Retroviral gene vector and expression of GFP in CD4+ T lymphocytes. a, Retroviral vector construct pLGFPSN used to transduce CD4+ T lymphocytes. 5′ LTR, long terminal repeat sequences (5′) driving expression of the transgene; SD and SA, splice donor and splice acceptor sequences; SV40, simian virus 40 promoter driving neomycin resistance gene NEO; 3′ LTR, long terminal repeat sequences (3′) containing polyadenylation sequence. b, Phase contrast (left) and fluorescence microscopy of MBP-specific T-lymphocyte blasts after transduction with GFP (green, middle panel). The cells were spread over an object slide and stained with a monoclonal mouse antibody against rat-CD4 (red, right panel). PE-labeled goat anti-mouse serum was used as secondary antibody. c, Activation-dependent GFP expression of CD4+ T lymphocytes. Histograms and dot plots of activated (upper) and resting (lower) transduced and non-transduced T lymphocytes. Dot plots show GFP-transduced T cells stained for CD4. A representative cell line of more than 15 tested is shown.